Performance Evaluation of a Commercial Real-Time PCR Method for the Detection of Lupin Traces in Food
Clara Tramuta,
Lucia Decastelli,
Francesco Ingravalle,
Elisa Barcucci,
Sandra Fragassi,
Daniela Manila Bianchi
Affiliations
Clara Tramuta
Experimental Zooprophylactic Institute of Piedmont, Liguria and Valle d’Aosta, Italian National Reference Center for the Detection of Food Allergens and Substances Causing Food Intolerance (CReNaRiA), 10154 Turin, Italy
Lucia Decastelli
Experimental Zooprophylactic Institute of Piedmont, Liguria and Valle d’Aosta, Italian National Reference Center for the Detection of Food Allergens and Substances Causing Food Intolerance (CReNaRiA), 10154 Turin, Italy
Francesco Ingravalle
Experimental Zooprophylactic Institute of Piedmont, Liguria and Valle d’Aosta, Department Biostatistics, Epidemiology and Risk Analysis (BEAR), 10154 Turin, Italy
Elisa Barcucci
Experimental Zooprophylactic Institute of Piedmont, Liguria and Valle d’Aosta, Italian National Reference Center for the Detection of Food Allergens and Substances Causing Food Intolerance (CReNaRiA), 10154 Turin, Italy
Sandra Fragassi
Experimental Zooprophylactic Institute of Piedmont, Liguria and Valle d’Aosta, Italian National Reference Center for the Detection of Food Allergens and Substances Causing Food Intolerance (CReNaRiA), 10154 Turin, Italy
Daniela Manila Bianchi
Experimental Zooprophylactic Institute of Piedmont, Liguria and Valle d’Aosta, Italian National Reference Center for the Detection of Food Allergens and Substances Causing Food Intolerance (CReNaRiA), 10154 Turin, Italy
In accordance with U.S. FDA Foods Program Regulatory Science Steering Committee guidelines, with this study, we optimized and validated a commercial real-time PCR method for the detection of low amounts of lupin in four classes of food matrices: chocolate cookies, ragù, Olivier salad, and barley and rice flour. DNA extracted from blank (true negative) samples artificially contaminated with lupin (Lupinus albus) flour at 1000 ppm underwent dilutions with the DNA extracted from the true negative samples up to 0.5 ppm. The limit of detection for real-time PCR was 0.5 ppm in the complex matrices (range, Ct 26–34), making this a specific, robust, and rapid method for lupin allergen detection and labeling. Our validation data support the suitability of this commercially available real-time PCR method for this purpose.
